目的：观察阿米洛利在经典的5/6肾切除大鼠模型中对蛋白尿的影响，探讨其影响蛋白尿的机制。方法构建经典的5/6肾切除大鼠模型，实验分为3组：假手术对照组（Sham）、5/6肾切除组（NTX）、5/6肾切除+阿米洛利干预组（NTX+阿米洛利）。采用考马斯亮蓝法检测各组24 h尿蛋白，利用免疫荧光共聚焦、实时荧光定量PCR等技术，检测足细胞尿激酶受体（uPAR）蛋白和uPAR mRNA的表达情况。结果第2周时，与假手术对照组（Sham）相比较，5/6肾切除组（NTX）24 h尿蛋白明显升高（47.50±28.05 mg vs 14.28±3.8 mg，P=0.023），有统计学意义；与NTX组相比，NTX+阿米洛利24 h尿蛋白无统计学意义（51.56±21.03 mg vs 47.50±28.05 mg，P=0.748）。第12周时，与NTX组（188.31±29.82）mg相比，NTX+阿米洛利组（121.37±31.14）mg、Sham组（21.32±8.59）mg 24 h尿蛋白增加量均进一步减少，具有统计学意义[P值分别为0.001，0.000]。激光共聚焦显微镜观察，与Sham组相比较，NTX组PLAUR mRNA表达明显增高（9.74±1.44 vs 1.01±0.13，P<0.01），有统计学意义；与NTX组相比较， NTX+阿米洛利mRNA表达明显降低（9.74±1.44 vs 5.01±1.36，P<0.01），有统计学意义。结论阿米洛利可能通过抑制受损足细胞uPAR的表达，从而起到降蛋白尿、延缓肾小球硬化的作用。%Objective To observe the effect of amiloride on the proteinuria of the 5/6 nephrectomy rats. Methods To establish the 5/6 nephrectomy rats model and divide the experiment into 3 groups, sham operated group(Sham), 5/6 nephrectomy model group(NTX) and 5/6 nephrectomy with amiloride-treated group (NTX+amiloride, n=15). The concentration of protein and mRNA of uPAR and the change of podocytes motility were detected by coomassiebluestaining, immunofluorence method and real-time PCR. Results At second week, compared with Control group, the 24 h urine protein of NTX group was significantly increased (47.50 ± 28.05 mg vs 14.28 ± 3.8 mg, P=0.023). There was no statistical significance in 24-hour urine protein between NTX+amiloride group and NTX group (51.56 ± 21.03 mg vs 47.50 ± 28.05 mg, P=0.748). The same situation was also observed at the time point of 12 week, comparing with NTX group, 24-hour urine protein decreased in Sham group (188.31 ± 29.82 mg vs 21.32±8.59 mg, P=0.000) and NTX+amiloride group (188.31±29.82 mg vs 121.37±31.14 mg, P=0.000), with statistical significance when comparing with Sham group, the expression of uPAR mRNA in NTX group was significantly increased (9.74 ± 1.44 vs 1.01±0.13, P=0.000). In contrast, the expression of uPAR mRNA in NTX rats treated with amiloride was significantly lower than in NTX group (9.74±1.44 vs 5.01±1.36, P=0.000). Conclusion Amiloride can reduce the proteinuria of the 5/6 nephrectomy rats model of transient proteinuria by inhibiting the induction of uPAR expression.
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